KR102102868B1 - A composition for preventing or treating neurodegenerative diseases comprising gossypetin - Google Patents

Abstract

The present invention relates to a composition for preventing or treating degenerative brain disease, comprising gossypetin or a salt thereof.
Since the composition of the present invention has an excellent effect on the prevention and treatment of degenerative brain disease, and the effect of improving memory and cognitive function is also excellent, the composition of the present invention is useful for preventing and treating degenerative brain disease, as well as improving memory and cognitive function Can be used.

Description

A composition for preventing or treating neurodegenerative diseases comprising gossypetin}

The present invention relates to a composition for preventing or treating degenerative brain disease and a method for treating degenerative brain disease using the same, comprising gossypetin or a salt thereof. In addition, the present invention relates to a composition for improving memory and cognitive function, comprising gosipetin or a salt thereof.

Due to the development of income level and medical and health environment, the proportion of the elderly population is gradually increasing, mainly in western countries. As of 2017, the proportion of the population aged 65 and over reached 14% and became an aging society. In 2025, it is expected to enter an aging society. Therefore, the aging disease of the elderly population is emerging as an urgent social problem.

In particular, Alzheimer's disease, which accounts for about half of senile dementia disease, currently has no effective causative agent and no prevention. There are five types of drugs prescribed by the Food and Drug Administration (FDA) that are prescribed to patients with Alzheimer's disease, including acetylcholine esterase inhibitors and glutamic acid receptor inhibitors, but they all only relieve symptoms.

Degenerative brain disease is caused by aggregation of proteins in brain tissue. In Alzheimer's disease, Aβ peptides are entangled to form plaques outside the cells. As a result, free radicals are generated and an inflammatory reaction caused by microglia occurs. IL-1, TNFα, PGE2, NO, NOO-, O ₂ ‥, H ₂ O _2, etc. are secreted from microglia to promote the death of surrounding neurons. Active oxygen species are caused by oxidative stress, and in the mitochondria, cytochrome C releases and activation of caspase-3 causes apoptosis. In addition, aggregation of Aβ peptides is increased, and aggregation of Tau proteins that stabilize microtubules present in neurons is also induced. When the microtubules collapse, the axons and dendrites of the nerve cells are atrophied, causing the degeneration of the brain nerve cells, and the tau protein aggregates and accumulates in the cells. do.

In addition to Alzheimer's disease, Huntington's disease causes aggregation of a protein called huntingtin, which accumulates in cells and causes cell death. In particular, when the CAG repeat sequence is genetically generated in the huntingtin gene, more than 100 glutamines are attached to the N-terminal of the protein, making it difficult to form a normal protein structure. In addition, when the function of chaperone protein is weakened, it is difficult to form a three-dimensional structure of the protein, and proteins that have misfolding are entangled with each other to form aggregates. In addition, in Parkinson's disease, aggregation of alpha-synuclein protein increases, forming a thread-like structure, and having an amyloid fiber structure. Then, the formation of Lewy bodies in cells increases, inhibiting the function of cells, and eventually causing neuronal cell death. Recently, the role of normal α-synuclein has been investigated. When the concentration of calcium ions in neurons increases, α-synuclein attaches to the endoplasmic reticulum that stores the neurotransmitter and secretion occurs normally. It acts as a calcium sensor, and a delicate balance is established between calcium and α-synuclein in the cell. Therefore, it is explained that when this balance is broken, aggregation of α-synuclein begins and leads to Parkinson's disease.

As such, a representative symptom of degenerative brain disease appears as an increase in protein aggregates in brain tissue. Due to the aggregation of proteins, an inflammatory immune response occurs in brain tissue and spreads to surrounding tissues. In addition, the function of the organelles in the cell is broken and the apoptosis reaction progresses, causing degeneration of brain tissue.

For degenerative brain disease, long-term use of the drug is required, so it is not very toxic and it is essential to develop a substance with good efficacy. Therefore, side effects can be minimized by developing substances that can be treated and prevented from natural products used for food.

On the other hand, Korean Patent Registration No. 10-1424547 and Korean Patent Publication No. 10-2015-0047687 disclose compositions for treating degenerative brain diseases, including products fermented by lactic acid bacteria or lactic acid bacteria, but they are degenerative such as dementia, etc. Since the therapeutic effect of brain disease is not high, there is a limit to commercial application.

Under these backgrounds, the present inventors have completed the present invention by confirming that the small molecule substance derived from natural products, gossypetin, is excellent in the prevention and treatment efficacy of degenerative brain disease after careful efforts.

An object of the present invention is to provide a pharmaceutical composition for preventing or treating degenerative brain disease, comprising gossypetin or a salt thereof.

Another object of the present invention is to provide a composition for inhibiting VRK2 activity, comprising gosipetin or a salt thereof.

Another object of the present invention is to provide a health functional food composition for the prevention or improvement of degenerative brain disease, including gosifetin or a salt thereof.

Another object of the present invention is to provide a pharmaceutical composition for improving memory or cognitive function comprising gosipetin or a salt thereof.

Another object of the present invention is to provide a health functional food composition for improving memory or cognitive function comprising gosifetin or a salt thereof.

Another object of the present invention is to provide a method for preventing or treating degenerative brain disease, comprising administering a composition comprising gosipine or a salt thereof to an individual other than a human.

Specifically, it is as follows. Meanwhile, each description and embodiment disclosed in the present invention can be applied to each other description and embodiment. That is, all combinations of the various elements disclosed in the present invention fall within the scope of the present invention. In addition, the scope of the present invention is not limited by the specific descriptions described below.

In addition, one of ordinary skill in the art can recognize or identify a number of equivalents to certain aspects of the invention described herein using only routine experimentation. In addition, such equivalents are intended to be included in the present invention.

One aspect of the present invention provides a pharmaceutical composition for preventing or treating degenerative brain disease, comprising Gossypetin or a salt thereof.

Another aspect of the present invention provides a health functional food composition for improving memory or cognitive function, comprising gosipetin or a salt thereof.

The composition may include gosifetin as an active ingredient.

In the present invention, "gosifetin" may be a compound represented by Formula 1 below:

[Formula 1]

The gosipetin is 2- (3,4-dihydroxyphenyl) -3,5,7,8-tetrahydroxy-4H-chromen-4-one (2- (3,4-dihydroxyphenyl) -3 , 5,7,8-tetrahydroxy-4H-chromen-4-one).

The gosifetin may be chemically synthesized, extracted from a plant, or a commercially available material, but the method for obtaining it is not particularly limited, and a method known in the art may be used.

The pharmaceutical composition of the present invention may include not only the above-described gosifetin, but also pharmaceutically acceptable salts thereof. In the present invention, "a pharmaceutically acceptable salt" means that the compound is in the form of a salt combined with another substance, and is capable of exhibiting similar pharmacological activity.

The types of pharmaceutically acceptable salts include, but are not limited to, inorganic acid salts such as hydrochloride, hydrobromide, phosphate or sulfate, and organic acid salts such as carboxylic acid salts or sulfonic acid salts. In addition, the type of carboxylate includes, but is not limited to, acetate, maleate, fumarate, malate, citrate, tartrate, lactate or benzoate. Further, the type of sulfonate includes, but is not limited to, methanesulfonate, ethanesulfonate, benzenesulfonate, toluene sulfonate, or naphthalenedisulfonate.

In the present invention, "degenerative brain disease" refers to a disease caused by brain cell damage.

The degenerative brain disease is caused by at least one of aggregation of protein, increased VRK2 activity, increased decomposition of chaperone protein, abnormal folding of protein, leusosome formation, polyglutamine aggregation, inflammatory response of glial cells, or pathology thereof. It may be shown in the red aspect, the protein aggregation, amyloid Aβ, tau protein, huntingtin (huntingtin), may be aggregation of α-synuclein, but is not limited thereto.

Specifically, the degenerative brain diseases of the present invention include dementia, Alzheimer's disease, Huntington's disease, Parkinson's disease, multiple system atrophy, multiple sclerosis, tauopathies, brain tumors, and peak disease Or it may be one or more diseases selected from the group consisting of Creutzfeldt-Jakob disease, and more specifically, one or more diseases selected from Alzheimer's disease, Huntington's disease, and Parkinson's disease, but is not limited thereto.

In the present invention, "memory / cognitive function" is an ability to efficiently manipulate knowledge and information, and includes all processes of thinking, speaking, remembering, judging, and executing using the brain. Memory and cognitive function may be lost or impaired due to brain cell damage, which may be manifested as symptoms such as forgetfulness, memory loss, and memory impairment.

The term "prevention" in the present invention means all actions to suppress or delay the onset of cognitive dysfunction or neuroinflammation by administration of the composition according to the present invention, and "treatment" is recognized by administration of the pharmaceutical composition. Suspicion of dysfunction or nerve inflammation and all actions in which the symptoms of an affected individual are improved or beneficially altered, and “improvement” refers to parameters related to the condition being treated with administration of the composition of the present invention, such as at least the severity of symptoms It means all acts of reduction.

In one embodiment of the present invention, it was confirmed that gosifetin inhibits the activity of VRK2 in vitro, and it was confirmed that by administering gosifetin to dementia-induced mice in vivo, memory is improved and dementia progression is inhibited. Therefore, the composition of the present invention can be used to inhibit VRK2 activity, and can be usefully used to improve memory and cognitive functions, as well as prevent and treat degenerative brain diseases.

The composition of the present invention may further include suitable carriers, excipients and diluents commonly used in the manufacture of pharmaceutical compositions. In addition, it can be formulated and used in the form of oral formulations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories, and sterile injectable solutions according to a conventional method.

Pharmaceutical compositions comprising gosipetin or a pharmaceutically acceptable salt thereof according to the present invention, external preparations and sterile injections, such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, respectively, according to a conventional method It can be formulated in the form of a solution, and may have a cream, gel, patch, spray, ointment, warning, lotion, linement, pasta, or cataplasma formulation, but is not limited thereto.

Carriers, excipients, and diluents that may be included in the composition comprising gosifetin include lactose, dextrose, sucrose, oligosaccharides, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium Phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. In the case of formulation, it is prepared using diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents and surfactants. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc. These solid preparations include at least one excipient such as starch, calcium carbonate, sucrose in the extract. ) Or by mixing lactose, gelatin, and the like. In addition, lubricants such as magnesium stearate and talc are used in addition to simple excipients. Liquid preparations for oral use include suspensions, liquid solutions, emulsions, syrups, etc. In addition to water and liquid paraffin, which are commonly used as diluents, various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, can be included. . Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solvents and suspensions may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. As a base for suppositories, witepsol, macrogol, tween 61, cacao butter, laurin butter, and glycerogelatin may be used.

Another aspect of the present invention provides a health functional food composition for preventing or improving degenerative brain disease, comprising gosipetin or a salt thereof.

Another aspect of the present invention provides a health functional food composition for improving memory or cognitive function, comprising gosipetin or a salt thereof.

The composition may include gosifetin as an active ingredient.

Gosifetin, degenerative brain disease, memory and cognitive function, prevention, treatment, and improvement are as described above.

When using the gosifetin of the present invention as a food additive, the gosifetin can be added as it is or used with other foods or food ingredients, and can be suitably used according to a conventional method. The mixing amount of the active ingredient can be appropriately determined according to the purpose of use (prevention, health or therapeutic treatment). Generally, when producing food or beverage, the gosipine of the present invention is added in an amount of 15% by weight or less, preferably 10% by weight or less, based on the raw material. However, in the case of long-term intake for health and hygiene purposes or for health control, the amount may be below the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount above the above range.

There are no particular restrictions on the type of food. Examples of foods to which the above substances can be added are meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other dairy products including noodle, gum, ice cream, various soups, beverages, teas, drinks, Alcoholic beverages and vitamin complexes, and all of the health functional foods in the ordinary sense.

The health drink composition according to the present invention may contain various flavoring agents, natural carbohydrates, and the like, as additional components, like a conventional beverage. The natural carbohydrates described above are monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol. As the sweetener, natural sweeteners such as taumatin and stevia extract, synthetic sweeteners such as saccharin and aspartame can be used. The proportion of the natural carbohydrate is generally about 0.01-0.20 g, preferably about 0.04-0.10 g per 100 mL of the composition of the present invention.

In addition to the above, the composition of the present invention includes various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, And carbonated agents used in carbonated beverages. In addition, the composition of the present invention may contain flesh for the preparation of natural fruit juice, fruit juice beverages and vegetable beverages. These ingredients can be used independently or in combination. The proportion of these additives is not critical, but is generally selected from 0.01-0.20 parts by weight per 100 parts by weight of the composition of the present invention.

Other ingredients that may be added include fats and oils, moisturizers, emollients, surfactants, organic and inorganic pigments, organic powders, ultraviolet absorbers, preservatives, fungicides, antioxidants, plant extracts, pH adjusters, alcohols, pigments, flavors, And blood circulation accelerators, cooling agents, restriction agents, and purified water.

Examples of the fat or oil component include ester fats and fats, hydrocarbon fats and fats, silicone fats and fats, fluorine fats and fats, animal fats, and vegetable fats and oils.

Examples of the ester fats and oils include glyceryl tri2-ethylhexanoate, cetyl 2-ethylhexanoate, isopropyl myristate, butyl myristate, isopropyl palmitate, ethyl stearate, octyl palmitate, isocetyl isostearate, and stearic acid. Butyl, ethyl linoleate, isopropyl linoleate, ethyl oleate, isocetyl myristate, isostearyl myristate, isostearyl palmitate, octyldodecyl myristate, isocetyl isostearate, diethyl sebacate, adipine Diisopropyl acid, isoalkyl neopentanoate, tri (capryl, capric acid) glyceryl, tri2-ethylhexanoate trimethylolpropane, triisostearate trimethylolpropane, tetra2-ethylhexanoate pentaerythritol , Cetyl caprylate, decyl laurate, hexyl laurate, decyl myristate, myristyl myristate, cetyl myristate, stearyl stearate, decyl oleate, ricinooleic acid , Isostearyl laurate, isotridecyl myristate, isocetyl palmitate, octyl stearate, isocetyl stearate, isodecyl oleate, octyldodecyl oleate, octyldodecyl oleate, isopropyl isoleate, 2 -Seto stearyl ethyl hexanoate, stearyl 2-ethyl hexanoate, hexyl isostearate, ethylene glycol dioctanoate, ethylene glycol dioleate, propylene glycol dicaprate, propylene glycol di (caprylic, capric acid) propylene glycol, dica Propyl glycol phthalate, neopentyl glycol dicapric acid, neopentyl glycol dioctanoate, glyceryl tricaprylic acid, glyceryl triundecylate, glyceryl triisopalmitate, glyceryl triisostearate, octyl dodecyl neopentane , Isostearyl octanoate, octyl isononanoate, hexyldecyl neodecanoate, octyldodecyl neodecanoate, isocetyl isostearate, isostearyl isostearate, Octyldecyl stearate, polyglycerin oleic acid ester, polyglycerin isostearic acid ester, triisocetyl citrate, triisoalkyl citrate, triisooctyl citrate, lauryl lactate, myristyl lactate, cetyl lactate, octyldecyl lactate, trityl citrate Ethyl, acetyl triethyl citrate, acetyl tributyl citrate, trioctyl citrate, diisostearyl malate, 2-ethylhexyl hydroxystearic acid, di2-ethylhexyl succinate, diisobutyl succinate, diisopropyl sebacate, Dioctyl sebacate, cholesteryl stearate, cholesteryl isostearate, cholesteryl hydroxystearate, cholesteryl oleate, dihydrocholesteryl oleate, phytsteryl isostearate, phytyl oleate, 12-stealoylhydroxystearate isocetyl, 12-stealoylhydroxystearate stearyl, 12-stealo And ester systems such as isostearyl monohydroxystearate.

Examples of hydrocarbon-based fats and oils include hydrocarbon-based fats and oils such as squalene, liquid paraffin, alpha-olefin oligomer, isoparaffin, ceresin, paraffin, liquid isoparaffin, polybutene, microcrystalline wax, and petrolatum.

Examples of silicone oils and fats include polymethylsilicone, methylphenylsilicone, methylcyclopolysiloxane, octamethylpolysiloxane, decamethylpolysiloxane, dodecamethylcyclosiloxane, dimethylsiloxane / methylcetyloxysiloxane copolymer, dimethylsiloxane / methylstearoxylsiloxane copolymer, and alkyl And modified silicone oils and amino modified silicone oils.

Perfluoropolyether etc. are mentioned as a fluorine-type oil and fat.

As animal or vegetable oil, avocado oil, almond oil, olive oil, sesame oil, rice bran oil, new flower oil, soybean oil, corn oil, rapeseed oil, almond oil, palm kernel oil, palm oil, castor oil, sunflower oil, grape seed oil , Cottonseed Oil, Palm Oil, Cucumin Nut Oil, Wheat Germ Oil, Rice Germ Oil, Shea Butter, Colostrum Colostrum, Marker Demi Nut Oil, Meadow Home Oil, Yolk Yolk Oil, Uji, Horse Oil, Mink Oil, Orange Rape Oil, Jojoba Oil , Candeler wax, carnauba wax, liquid lanolin, hardened castor oil and other animal or plant fats.

Examples of the moisturizing agent include water-soluble low molecular moisturizing agents, oil-soluble molecular moisturizing agents, water-soluble polymers, and fat-soluble polymers.

Examples of water-soluble low-molecular moisturizers include serine, glutamine, sorbitol, mannitol, sodium pyrrolidone-sodium carboxylate, glycerin, propylene glycol, 1,3-butylene glycol, ethylene glycol, polyethylene glycol B (polymerization degree n = 2 or higher), and polypropylene. Glycol (polymerization degree n = 2 or more), polyglycerin B (polymerization degree n = 2 or more), lactic acid, lactate, and the like.

Examples of fat-soluble low-molecular moisturizing agents include cholesterol and cholesterol esters.

Water-soluble polymers include carboxyvinyl polymer, polyasparaginate, tragacanth, xanthan gum, methylcellulose, hydroxymethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, carboxymethylcellulose, water-soluble chitin, chitosan, dextrin, etc. You can.

Examples of the fat-soluble polymer include polyvinylpyrrolidone-eicocene copolymer, polyvinylpyrrolidone-hexadecene copolymer, nitrocellulose, dextrin fatty acid ester, and polymer silicone.

Examples of the emollient agent include long-chain acyl glutamate cholesteryl ester, hydroxystearate cholesteryl, 12-hydroxystearic acid, stearic acid, rosin acid, lanolin fatty acid cholesteryl ester, and the like.

Examples of the surfactant include nonionic surfactants, anionic surfactants, cationic surfactants and amphoteric surfactants.

As nonionic surfactants, self-emulsifying glycerin monostearate, propylene glycol fatty acid ester, glycerin fatty acid ester, polyglycerin fatty acid ester, sorbitan fatty acid ester, POE (polyoxyethylene) sorbitan fatty acid ester, POE sorbit fatty acid ester, POE Glycerin fatty acid ester, POE alkyl ether, POE fatty acid ester, POE hardened castor oil, POE castor oil, POE ㆍ POP (polyoxyethylene ㆍ polyoxypropylene) copolymer, POE ㆍ POP alkyl ether, polyether-modified silicone, lauric acid And alkanolamides, alkylamine oxides, and hydrogenated soybean phospholipids.

Examples of the anionic surfactant include fatty acid soap, alpha-acyl sulfonate, alkyl sulfonate, alkyl allyl sulfonate, alkyl naphthalene sulfonate, alkyl sulfate, POE alkyl ether sulfate, alkylamide sulfate, alkyl phosphate, POE alkyl phosphate, and alkylamide phosphate. , Alkyloyl alkyl taurine salts, N-acyl amino acid salts, POE alkyl ether carboxylate salts, alkyl sulfosuccinate salts, sodium alkyl sulfoacetate salts, acylated hydrolyzed collagen peptide salts, perfluoroalkyl phosphate esters, and the like. .

Examples of the cationic surfactant include alkyltrimethylammonium chloride, stearyltrimethylammonium chloride, stearyltrimethylammonium bromide, cetostearyltrimethylammonium chloride, distearyldimethylammonium chloride, stearyldimethylbenzylammonium chloride, behenyltrimethylammonium bromide, and chloride. And benzalkonium, diethylaminoethyl stearate, dimethylaminopropylamide stearate, and quaternary ammonium salts of lanolin derivatives.

Examples of the amphoteric surfactant include carboxybetaine type, amidebetaine type, sulfobetaine type, hydroxysulfobetaine type, amidesulfobetaine type, phosphobetaine type, aminocarboxylate type, imidazoline derivative type, and amideamine type. And amphoteric surfactants.

Examples of organic and inorganic pigments include silicic acid, silicic anhydride, magnesium silicate, talc, sericite, mica, kaolin, bengala, clay, bentonite, titanium film mica, bismuth oxychloride, zirconium oxide, magnesium oxide, zinc oxide, titanium oxide, and aluminum oxide , Inorganic pigments such as calcium sulfate, barium sulfate, magnesium sulfate, calcium carbonate, magnesium carbonate, iron oxide, ultramarine, chromium oxide, chromium hydroxide, calamine and complexes thereof; Polyamide, polyester, polypropylene, polystyrene, polyurethane, vinyl resin, urea resin, phenol resin, fluorine resin, silicon resin, acrylic resin, melamine resin, epoxy resin, polycarbonate resin, divinylbenzene ㆍ styrene copolymer, And organic pigments such as silk powder, cellulose, CI pigment yellow, and CI pigment orange, and composite pigments of these inorganic and organic pigments.

Examples of the organic powder include metal soaps such as calcium stearate; Alkyl phosphate metal salts such as sodium cetyl acid zinc, lauryl acid zinc, and calcium lauryl acid; Acylamino acid polyvalent metal salts such as N-lauroyl-beta-alanine calcium, N-lauroyl-beta-alanine zinc, and N-lauroylglycine calcium; Amide sulfonic acid polyvalent metal salts such as N-lauroyl-taurine calcium and N-palmitoyl-taurine calcium; N such as N-epsilon-lauroyl-L-lysine, N-epsilon-palmitoylazine, N-alpha-paritoylolnitine, N-alpha-lauroylarginine, N-alpha-cured Uji fatty acid acyl arginine -Acyl basic amino acids; N-acylpolypeptides, such as N-lauroylglycylglycine; Alpha-amino fatty acids such as alpha-amino caprylic acid and alpha-amino lauric acid; And polyethylene, polypropylene, nylon, polymethyl methacrylate, polystyrene, divinylbenzene-styrene copolymer, and ethylene tetrafluoride.

Examples of ultraviolet absorbers include paraaminobenzoic acid, ethyl paraaminobenzoate, amyl paraaminobenzoate, octyl paraaminobenzoate, ethylene glycol salicylate, phenyl salicylate, octyl salicylate, benzyl salicylate, butyl salicylate, homomentyl salicylate, benzyl cinnamate , Paramethoxy cinnamic acid-2-ethoxyethyl, paramethoxy cinnamic acid octyl, diparamethoxy cinnamic acid mono-2-ethylhexaneglyceryl, paramethoxy cinnamic acid isopropyl, diisopropyl-diisopropyl cinnamic acid ester mixture, uro Canonic acid, ethyl urocanate, hydroxymethoxybenzophenone, hydroxymethoxybenzophenone sulfonic acid and salts thereof, dihydroxymethoxybenzophenone, dihydroxymethoxybenzophenone disulfonate sodium, dihydroxybenzophenone , Tetrahydroxybenzophenone, 4-tert-butyl-4'-methoxydibenzoylmethane, 2,4,6-trianilino-p- (carbo-2'-ethylhexyl-1'-oxy) -1 , 3,5-triazine, 2- (2-hydroxy Ci-5-methylphenyl) benzotriazole etc. are mentioned.

As a fungicide, hinokitiol, triclosan, trichlorohydroxydiphenyl ether, chlorhexidine gluconate, phenoxyethanol, resorcin, isopropylmethylphenol, azulene, salicylic acid, zincfilionone, benzalkonium chloride, photosensitive No. 301, sodium mononitrowire, and undeciric acid.

Examples of antioxidants include butyl hydroxyanisole, propyl gallic acid, and elisoric acid.

Examples of the pH adjusting agent include citric acid, sodium citrate, malic acid, sodium malate, phthalic acid, sodium phthalate, succinic acid, sodium succinate, sodium hydroxide, and sodium monohydrogen phosphate.

Examples of the alcohol include higher alcohols such as ethyl alcohol.

In addition, the compounding component that may be added in addition is not limited to this, and any of the above components can be blended within a range not impairing the objects and effects of the present invention, but preferably 0.01-5% by weight based on the total weight, More preferably, it is blended at 0.01-3% by weight.

When the formulation of the present invention is a lotion, paste, cream or gel, animal fibers, plant fibers, wax, paraffin, starch, trakant, cellulose derivatives, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide as carrier components Can be used.

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder can be used as a carrier component, and in the case of a spray, additionally chlorofluorohydrocarbon, propane / Propellant such as butane or dimethyl ether.

When the formulation of the present invention is a solution or emulsion, a solvent, solvating agent or emulsifying agent is used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 , 3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or fatty acid ester of sorbitan.

When the formulation of the present invention is a suspension, liquid diluents such as water, ethanol or propylene glycol as carrier components, suspensions such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, microcrystalline Cellulose, aluminum metahydroxide, bentonite, agar or trakant, etc. can be used.

When the formulation of the present invention is a surfactant-containing cleansing, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivatives, methyltaurate, sarcosinate, fatty acid amide as a carrier component Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, linoline derivatives or ethoxylated glycerol fatty acid esters and the like can be used.

Another aspect of the present invention provides a method for preventing or treating degenerative brain disease, comprising administering the pharmaceutical composition to an individual.

In the present invention, "individual" means a subject in need of treatment of a disease. Specifically, it may mean a mammal such as a human or non-human primate, a mouse, a rat, a dog, a cat, a horse, and a cow, and more specifically an animal other than a human, Or it may mean a human, but is not limited thereto.

The pharmaceutical composition according to the present invention can be administered in a pharmaceutically effective amount. The "pharmaceutically effective amount" means an amount sufficient to treat the disease at a reasonable benefit / risk ratio applicable to medical treatment, and the effective dose level is the type, severity, activity of the drug, and sensitivity to the drug. , Time of administration, route of administration and rate of discharge, duration of treatment, factors including co-drugs and other factors well known in the medical field. In one embodiment, the concentration of gosipetine administered may be 0.1 mg / kg or more and 500 mg / kg or less, and specifically 0.1 mg / kg or more and 100 mg / kg or less, 0.5 mg / kg or more and 100 mg / g, based on the weight of the administered individual body. kg or less, 0.5mg / kg or more, 50mg / kg or less, 0.5mg / kg or more, 30mg / kg or less, 0.5mg / kg or more, 15mg / kg or less, 7mg / kg or more, 13mg / kg or less, more specifically 10mg / kg concentration Gossifetin may be administered, but is not limited thereto. The pharmaceutical composition according to the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with a conventional therapeutic agent, and may be administered single or multiple. In consideration of all the above factors, it is important to administer an amount capable of obtaining the maximum effect in a minimal amount without side effects, which can be easily determined by those skilled in the art to which the present invention pertains.

The pharmaceutical composition of the present invention can be administered to a subject by various routes. All modes of administration can be expected, for example, by oral administration, subcutaneous, intravenous, intramuscular, intrauterine dura or intracranial injection. The pharmaceutical composition of the present invention is determined according to the type of drug that is the active ingredient along with various related factors such as the disease to be treated, the route of administration, the patient's age, sex, weight, and the severity of the disease.

The present invention has been found that the superior effect of inhibiting protein aggregation in brain cells, and in animal experiments, it has been found that the effect of preventing and treating and improving memory of degenerative brain disease is excellent. It can be useful for preventing and treating degenerative brain diseases and improving memory and cognitive functions.

Figure 1 is a result showing the protein aggregation inhibitory effect of gosifetin in cells. (a) shows the amount of protein aggregates that did not pass through the filter after treatment with gosifetin on the cells, and (b) was confirmed by electrophoresis of the amount of protein aggregates on the gel. (c) shows the formation of protein aggregates in cells and the reduction according to treatment with gosipine as agglutination of green fluorescent protein, and (d) is a quantitative analysis. (e) comparatively analyzed the amount of insoluble aggregates compared to soluble protein aggregates, and (f) analyzed the amount of water-soluble protein aggregates. (g) measured the number of cells with this protein aggregate, (h) is the result of measuring the total number of cells with the expression of the fluorescent protein.
Figure 2 is a result showing the degree of inhibition of VRK2 activity according to the concentration of gosifetin.
3 is a result of re-evaluating short-term memory of Alzheimer's induced mice through Y-shape labyrinth test for gosipine.
FIG. 4 shows the results of improving the learning function by measuring escape latency of Alzheimer's induced mice through the Morris underwater maze test for gosifetin.
5 is a result of measuring the memory capacity of the platform position of Alzheimer's induced mice through the Morris underwater maze test for gosifetin. (a) measures the time to stay where the platform was after removing the platform, and (b) measures the total distance of the mouse in the underwater maze test.
6 is a result of measuring the degree of decrease in the deposition of amyloid aggregates in the hippocampus of the mouse brain with respect to gosifetin.
7 is a result of biochemically measuring the degree of deposition of amyloid aggregates in mouse brain with respect to gosifetin. (a) shows the amount of amyloid aggregates that did not pass through the filter among the protein aggregates accumulated in the brain, and (b) measured the amount of soluble amyloid aggregates. (c) is an analysis of the amyloid aggregated polymer accumulated in the brain by electrophoresis, and (d) is a comparative analysis of the amount of the amyloid monolith and the polymer, respectively.
8 is a result of measuring the degree of reduction of gliosis by microglia in mouse brain for gosipine.
9 is a result of measuring the degree of reduction of gliosis by astrocytes in mouse brain with respect to gosifetin.
FIG. 10 shows the results of comparing the activity of inhibiting VRK2 activity with that of gosipetin for compounds known to significantly inhibit the aggregation of Aβ and Tau proteins in vitro.
FIG. 11 is a result of comparing the degree of inhibition of protein aggregation in cells with the efficacy of gosipine for compounds known to significantly inhibit the aggregation of Aβ and Tau proteins in vitro.
FIG. 12 is a result of comparing the degree of improvement of short-term memory capacity of Alzheimer's dementia-induced mice with the efficacy of gosipetin in a Y-maze experiment for compounds known to significantly inhibit the aggregation of Aβ and Tau proteins in vitro.
FIG. 13 shows the results of comparing the improvement of short-term memory capacity of Alzheimer's dementia-induced mice with the efficacy of gosipine in a Y-maze experiment for compounds known to significantly inhibit the aggregation of Aβ and Tau proteins in vitro.

Hereinafter, the present invention will be described in more detail through examples and experimental examples. However, these examples and experimental examples are for illustrative purposes only, and the scope of the present invention is not limited to these examples and experimental examples.

Example 1: Confirmation of in vitro effect of gosifetin

In vitro experiments were conducted to confirm that the natural product-derived compound, gosifetin, was effective in inhibiting degenerative brain disease.

Example 1-1: Confirmation of the effect of inhibiting aggregation of polyglutamine protein in cells that are concentration-dependent of gosipine

Agglutination of polyglutamine protein causes the development of degenerative brain disease. Dot blot assay method by expressing polyglutamine protein bound with green fluorescent protein in U2OS cells expressing VRK2 to check the effect of gosipine inhibiting aggregation of polyglutamine protein, and then treating gosipine by concentration It was confirmed by. In addition, the protein aggregates generated in the cells were observed by electrophoresis, and the degree of protein aggregate formation in the cells was observed with a fluorescent microscope and quantitatively displayed.

As a result, it was confirmed that the polyglutamine protein aggregate decreases depending on the concentration of gosipine (FIGS. 1A and 1B). In addition, it was confirmed that the insoluble protein aggregate was significantly reduced (FIG. 1C-E), and the number of cells with the protein aggregate was also decreased (FIG. 1F-1H).

Example 1-2: Confirmation of the cross-linking of gosifetin (GSP) and VRK2 and inhibition of the concentration-dependent VRK2 activity of gosifetin and the preservation of chaperone protein

VRK2, a phosphorylase, is known to regulate the TRiC chaperone protein in cells to prevent the removal of polyglutamine aggregates. Therefore, if VRK2 is inhibited, it is expected that polyglutamine aggregation can be suppressed, and in order to confirm whether gosifetin has an inhibitory effect on VRK2 activity, gosifetin was treated at various concentrations to measure VRK2 activity inhibition.

First, the combination of gosipine and sepharose 4B chemically (FIG. 2A), and then it was confirmed whether the gosipine and VRK2 bind to each other. VRK2 does not attach to Sepharose 4B, and it can be seen that VRK2 and gosifetin bind to each other by confirming that it binds only to gosifetin-sepharose 4B (Figure 2B). In addition, by confirming that the addition of gosifetin interferes with the binding between VRK2 and gosifetin-sepharose 4B, it was confirmed that the reciprocal linkage between gosifetin and VRK2 is specific (Figure 2C). And it showed that the activity of VRK2 was inhibited depending on the concentration of gosifetin (Fig. 2D), and it was confirmed that the amount of the chaperone protein, CCT-4, did not decrease with the addition of gosifetin (Fig. 2E).

As a result of the experiment, it was confirmed that gosifetin inhibits VRK2 activity in vitro, and thus was predicted to be effective in the prevention and treatment of degenerative brain disease.

Example 2: Confirmation of in vivo effect of gosifetin

Through the results of Example 1, an animal experiment was conducted to confirm whether the antioxidant brain disease is expected to be effective in treating and treating gosipine even in an actual animal.

In the experiment, a 5 week-old 5xFAD Alzheimer's-induced mouse was used, and gosipine was orally administered at a concentration of 10 mg / kg for 14 weeks (FIGS. 3A and 3B). Thereafter, a Y-shaped maze test and a Morris underwater maze experiment, which are widely used for cognitive function verification, were performed. Specifically, short-term memory was measured through the Y-shaped maze experiment. A week after the Y-shaped labyrinth experiment, a Morris underwater labyrinth experiment was conducted, and after 5 days, the mouse was trained to remember the location of the platform, and the time to visit the platform was measured. In addition, after removing the platform from the tank on the 6th day after the platform memory training, the frequency of the mouse staying in the section where the platform is located was measured.

As a result of the Y-shaped labyrinth experiment, it was confirmed that 5xFAD Alzheimer's induced mice administered with gosipetine improved short-term memory to a degree similar to that of normal mice (FIG. 3C). In addition, as a result of confirming that the Alzheimer's induced mouse remembers the platform through the Morris underwater maze experiment, it was confirmed that the gosipetine administration group finds the platform within a shorter time than the Alzheimer's mouse as well as the normal mouse (FIG. 4).

In addition, after removing the platform, as a result of confirming the frequency of staying in the section where the platform is located, the total distance moved is similar, but in the group fed with gosipetin to the Alzheimer's induced mouse, the frequency of staying in the section where the platform was located increased, so that the Alzheimer's induced mouse It was confirmed that the memory capacity was increased (FIG. 5).

Through this, it was proved that Gossifetin has an Alzheimer's prevention and treatment effect, and it was confirmed that it has a very excellent effect in improving cognitive function.

Example 3: Alzheimer's induced mouse brain tissue analysis treated with gosifetin

Example 3-1: Tissue analysis of Alzheimer's induced mouse brain treated with gosifetin

Alzheimer's-induced mice (5xFAD) were sectioned by extracting the brains of the group fed with gosipine and the group not fed, and then the degree of production of Aβ amyloid aggregates was analyzed by the method of staining Thioflavin S. When the hippocampus (Fig. 6A-6C) and frontal lobe (Fig. 6D-6F) tissues of the brain were observed, it was confirmed that Aβ amyloid aggregates were significantly reduced in the mouse group fed with gosipine.

Example 3-2: Biochemical analysis of the hippocampus region of Alzheimer's induced mouse brain treated with gosipetin

When the brain tissues of Alzheimer's-induced mice fed with gosipine were analyzed by biochemical analysis methods such as protein quantification and Western blot, both the amount of water-soluble Aβ amyloid protein in the dot blot and Western blot results were statistically significant It can be confirmed that the decrease (Fig. 7A-E). Aβ amyloid protein is a protein produced by the APP amyloid precursor protein being cut by an enzyme. When Western blot was conducted to confirm whether the decrease in Aβ amyloid protein was caused by a decrease in the amount of APP amyloid precursor protein, it was confirmed that the amount of APP amyloid precursor protein did not change (FIGS. 7F and 7G). .

Example 3-3: Analysis of degree of gliosis caused by hyperactivity of microglial cells in Alzheimer's induced mouse brain treated with gosipine

Alzheimer's-induced mice (5xFAD) were excised by excising the brains of the groups fed and not fed the gosipine group, and then GFAP, a specific protein of antibodies and astrocytes, a specific protein of microglia cells, IB-1 The distribution and proliferation of microglia and astrocytes were analyzed using Korean antibodies.

Hyperactivity of microglia and astrocytes in the mouse group fed with gosipetine, when the hippocampus of the brain (Fig. 8A-8B, Fig. 9A-9B) and frontal lobe (Fig. 8C-8D, Fig. 9C-9D) were observed, That is, it can be seen that gliosis is greatly reduced.

Example 4: Comparison of the inhibitory effect of gosifetin with other compounds on VRK2 activity

The level of inhibition of VRK2 activity of gosifetin was compared with other compounds known to inhibit aggregation of proteins that cause degenerative brain disease at the in vitro level.

Specifically, in vitro kinase assay measures VRK2 inhibitory activity against Rosmarinic acid and Lacmoid, which are known to significantly inhibit the aggregation of Aβ and Tau proteins, which are the causes of degenerative brain disease, at the in vitro level Did. In addition, by inducing polyglutamine protein aggregation to the cells, each compound was treated to observe the degree of reduction in protein aggregation in the cells with a fluorescence microscope. In addition, the total area of protein aggregation in cells, the average size of protein aggregation, and the degree of protein aggregation inhibition compared to the control group without drugs are shown in the table. Cytotoxicity of each compound was also confirmed.

As a result of measuring the autophosphorylation activity of VRK2, at the same concentration of 25 μM, gosipine most strongly inhibited the autophosphorylation activity of VRK2, and rosemary acid had a very weak inhibitory effect, and lacmoid had a slight inhibition of VRK2 activity. Showed (Fig. 10).

In addition, each compound showed little cytotoxicity (FIG. 11C), while gosipetin inhibited intracellular protein aggregation by 43%, while other compounds did not inhibit intracellular protein aggregation (FIGS. 11A-11B).

Through this, it was confirmed that the best effect of inhibiting protein aggregation without showing toxic toxicity in the cells. In addition, it was confirmed that even if there is an activity that inhibits some of the causative agent of Alzheimer's in vitro, that is, in vitro level, it is confirmed that it does not have activity against all of the causative agent of Alzheimer's. Did.

Example 5: Comparison of in vivo effects of gosifetin with other compounds

The compound of Example 4 was administered to the Alzheimer's mouse model to confirm the effect on actual cognitive function improvement.

Example 5-1: Alzheimer's prevention and treatment effect confirmed through Y maze experiment

Specifically, Alzheimer's-induced mouse model 5xFAD was used, and after 5 weeks of age of 5xFAD, Gossifetin (GSP), rosmarinic acid, and lacmoid were each orally administered at 10 mg / kg for 15 weeks, and then Y labyrinth experiment was performed. Cognitive function was tested.

As a result of the experiment, the performance of the 5xFAD mouse was significantly lower than that of the normal mouse (Fig. 12A), and a compound known to inhibit the aggregation of Aβ and Tau proteins in vitro, lacmoid (Fig. 12B) or rosmarinic acid (Fig. 12C) Treatment also did not contribute to improving this cognitive decline. However, it was confirmed that the administration of gosifetin restored the behavioral change level to almost normal level (FIG. 12D), and the cognitive function was improved according to the gosifetin administration.

Example 5-2: Alzheimer's prevention and treatment effect confirmed through an underwater maze experiment

Specifically, cognitive function was tested by orally administering gosipetin (GSP), rosmarinic acid, and racmoid at 10 mg / kg for 16 weeks from the time when the dementia-inducing mouse 5xFAD was 8 weeks old, and then performing an underwater maze experiment. The underwater maze experiment was conducted for 5 days after training to find the platform under the surface of the tank, and after removing the platform on the 6th day, it was confirmed how much the platform was remembered.

As a result of the experiment, the memory capacity of the 5xFAD mouse was significantly lower than that of the normal mouse (Fig. 13A). In addition, when treated with a compound known to inhibit the aggregation of Aβ and Tau proteins in vitro, racmoid and rosemarine, the cognitive function is lower than that when no administration was given (Fig. 13B), or that it is not able to recover significantly. It was confirmed (Fig. 13C). However, it was confirmed that the administration of gosifetin showed better memory ability than normal mice (Fig. 13D).

Through this, it was confirmed that even in the case of a compound known to be effective in vitro, the results of in vivo experiments were different, and it was confirmed that gosifetin had a very good effect on improving cognitive function.

From the above description, those skilled in the art to which the present invention pertains will understand that the present invention may be implemented in other specific forms without changing its technical spirit or essential characteristics. In this regard, it should be understood that the embodiments described above are illustrative in all respects and not restrictive. The scope of the present invention should be construed as including all changes or modifications derived from the meaning and scope of the following claims rather than the above detailed description and equivalent concepts thereof.

Claims (9)

A pharmaceutical composition for preventing or treating Alzheimer's, comprising Gossypetin or a salt thereof.
The pharmaceutical composition for preventing or treating Alzheimer's disease according to claim 1, wherein the gosipine is represented by Formula 1 below:
[Formula 1]

.
The pharmaceutical composition for preventing or treating Alzheimer's disease according to claim 1, wherein the Alzheimer's disease is caused by one or more causes selected from increased activity of VRK2, aggregation of beta amyloid (Aβ) protein, and aggregation of Tau protein.
delete delete delete A health functional food composition for preventing or improving Alzheimer's, comprising gosipetin or a salt thereof.
delete A method for preventing or treating Alzheimer's disease, comprising administering a composition comprising gosipine or a salt thereof to an individual other than a human.

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